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Article

Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer

  • Authors:
    • Lishui Wang
    • Yimin Liu
    • Lutao Du
    • Juan Li
    • Xiumei Jiang
    • Guixi Zheng
    • Ailin Qu
    • Haiyan Wang
    • Lili Wang
    • Xin Zhang
    • Hui Liu
    • Hongwei Pan
    • Yongmei Yang
    • Chuanxin Wang
  • View Affiliations / Copyright

    Affiliations: Department of Clinical Laboratory, Qilu Hospital, Shandong University, Jinan, Shandong 250012, P.R. China
  • Pages: 615-622
    |
    Published online on: March 4, 2015
       https://doi.org/10.3892/mmr.2015.3428
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Abstract

Serum microRNAs (miRNAs) have been proposed as novel non‑invasive biomarkers for the early detection of cancer. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) is the most commonly used method for investigating miRNA expression levels, however, the interpretation of RT‑qPCR results depends largely on normalization to an appropriate endogenous control. The present study involved 129 patients with non‑muscle‑invasive bladder cancer (NMIBC), 121 patients with muscle‑invasive bladder cancer (MIBC) and 158 healthy controls. The aim of the present study was to determine the most stable reference genes for the investigations of serum miRNA in bladder cancer (BC). MiSeq sequencing was performed and the expression levels of 10 miRNAs and U6 were then measured using RT‑qPCR. Following RT‑qPCR, five genes (hsa‑miR‑193a‑5p, hsa‑miR‑16‑5p, U6, hsa‑miR‑191‑5p and hsa‑let‑7d‑3p) were selected for stability analysis using geNorm and NormFinder software. These algorithms identified hsa‑miR‑193a‑5p and hsa‑miR‑16‑5p as the most stably expressed reference genes. The availability of hsa‑miR‑193a‑5p and hsa‑miR‑16‑5p was confirmed in an additional cohort. One‑way analysis of variance indicated that no significant differences were present in the expression levels among the three groups. Furthermore, miR‑148b‑3p was selected as a target miRNA to determine the effect of hsa‑miR‑193a‑5p and hsa‑miR‑16‑5p on miRNA quantification. The combined use of hsa‑miR‑193a‑5p and hsa‑miR‑16‑5p enabled the detection of a significant upregulation of miR‑148b‑3p in the BC serum. The results of the present study demonstrated that normalization of miRNA data, using a combination of hsa‑miR‑193a‑5p and hsa‑miR‑16‑5p as reference genes, may produce reliable and accurate results for the detection of serum miRNAs in BC.
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1 

Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar : PubMed/NCBI

2 

Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI

3 

Kaufman DS, Shipley WU and Feldman AS: Bladder cancer. Lancet. 374:239–249. 2009. View Article : Google Scholar : PubMed/NCBI

4 

Botteman MF, Pashos CL, Redaelli A, Laskin B and Hauser R: The health economics of bladder cancer: A comprehensive review of the published literature. Pharmacoeconomics. 21:1315–1330. 2003. View Article : Google Scholar

5 

Malmström PU: Why has the survival of patients with bladder cancer not improved? BJU Int. 101:267–269. 2008. View Article : Google Scholar

6 

Falebita OA, Sweeney P and Lee G: Urine cytology in the evaluation of urological malignancy revisited: is it still necessary. Urol Int. 84:45–49. 2010. View Article : Google Scholar

7 

Mengual L, Lozano JJ, Ingelmo-Torres M, Gazquez C, Ribal MJ and Alcaraz A: Using microRNA profiling in urine samples to develop a non-invasive test for bladder cancer. Int J Cancer. 133:2631–2641. 2013.PubMed/NCBI

8 

Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI

9 

Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI

10 

Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI

11 

Heneghan HM, Miller N and Kerin MJ: MiRNAs as biomarkers and therapeutic targets in cancer. Curr Opin Pharmacol. 10:543–550. 2010. View Article : Google Scholar : PubMed/NCBI

12 

Mitchell PS, Parkin RK, Kroh EM, et al: Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci USA. 105:10513–10518. 2008. View Article : Google Scholar : PubMed/NCBI

13 

Hunter MP, Ismail N, Zhang X, et al: Detection of microRNA expression in human peripheral blood microvesicles. PLoS ONE. 3:e36942008. View Article : Google Scholar : PubMed/NCBI

14 

Chim SS, Shing TK, Hung EC, Leung TY, Lau TK, Chiu RW and Lo YM: Detection and characterization of placental microRNAs in maternal plasma. Clin Chem. 54:482–490. 2008. View Article : Google Scholar : PubMed/NCBI

15 

Genovesi LA, Anderson D, Carter KW, Giles KM and Dallas PB: Identification of suitable endogenous control genes for microRNA expression profiling of childhood medulloblastoma and human neural stem cells. BMC Res Notes. 5:5072012. View Article : Google Scholar : PubMed/NCBI

16 

Chang KH, Mestdagh P, Vandesompele J, Kerin MJ and Miller N: MicroRNA expression profiling to identify and validate reference genes for relative quantification in colorectal cancer. BMC Cancer. 10:1732010. View Article : Google Scholar : PubMed/NCBI

17 

Zheng G, Wang H, Zhang X, et al: Identification and validation of reference genes for qPCR detection of serum microRNAs in colorectal adenocarcinoma patients. PLoS ONE. 8:e830252013. View Article : Google Scholar : PubMed/NCBI

18 

Cheng L, Montironi R, Davidson DD and Lopez-Beltran A: Staging and reporting of urothelial carcinoma of the urinary bladder. Mod Pathol. 22(Suppl 2): S70–S95. 2009. View Article : Google Scholar : PubMed/NCBI

19 

Asaga S, Kuo C, Nguyen T, Terpenning M, Giuliano AE and Hoon DS: Direct serum assay for microRNA-21 concentrations in early and advanced breast cancer. Clin Chem. 57:84–91. 2011. View Article : Google Scholar

20 

Zheng G, Wang H, Zhang X, Yang Y, Wang L, Du L, Li W, Li J, Qu A, Liu Y and Wang C: Identification and validation of reference genes for qPCR detection of serum microRNAs in colorectal adenocarcinoma patients. PLoS One. 8:e830252013. View Article : Google Scholar : PubMed/NCBI

21 

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A and Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3:research0034. 2002, View Article : Google Scholar : PubMed/NCBI

22 

Adam L, Wszolek MF, Liu CG, Jing W, Diao L, Zien A, Zhang JD, Jackson D and Dinney CP: Plasma microRNA profiles for bladder cancer detection. Urol Oncol. 31:1701–1708. 2013. View Article : Google Scholar

23 

Selth LA, Townley SL, Bert AG, Stricker PD, Sutherland PD, Horvath LG, Goodall GJ, Butler LM and Tilley WD: Circulating microRNAs predict biochemical recurrence in prostate cancer patients. Br J Cancer. 109:641–650. 2013. View Article : Google Scholar : PubMed/NCBI

24 

Köberle V, Kronenberger B, Pleli T, et al: Serum microRNA-1 and microRNA-122 are prognostic markers in patients with hepatocellular carcinoma. Eur J Cancer. 49:3442–3449. 2013. View Article : Google Scholar : PubMed/NCBI

25 

Huang X, Liang M, Dittmar R and Wang L: Extracellular microRNAs in urologic malignancies: Chances and challenges. Int J Mol Sci. 14:14785–14799. 2013. View Article : Google Scholar : PubMed/NCBI

26 

Liu R, Zhang C, Hu Z, et al: A five-microRNA signature identified from genome-wide serum microRNA expression profiling serves as a finger-print for gastric cancer diagnosis. Eur J Cancer. 47:784–791. 2010. View Article : Google Scholar

27 

Bediaga NG, Davies MP, Acha-Sagredo A, et al: A microRNA-based prediction algorithm for diagnosis of non-small lung cell carcinoma in minimal biopsy material. Br J Cancer. 109:2404–2411. 2013. View Article : Google Scholar : PubMed/NCBI

28 

Bargaje R, Hariharan M, Scaria V and Pillai B: Consensus miRNA expression profiles derived from interplatform normalization of microarray data. RNA. 16:16–25. 2010. View Article : Google Scholar :

29 

Chen X, Ba Y, Ma L, et al: Characterization of microRNAs in serum: A novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 18:997–1006. 2008. View Article : Google Scholar : PubMed/NCBI

30 

Pritchard CC, Kroh E, Wood B, Arroyo JD, Dougherty KJ, Miyaji MM, Tait JF and Tewari M: Blood cell origin of circulating microRNAs: A cautionary note for cancer biomarker studies. Cancer Prev Res (Phila). 5:492–497. 2012. View Article : Google Scholar

31 

Scheffer AR, Holdenrieder S, Kristiansen G, von Ruecker A, Müller SC and Ellinger J: Circulating microRNAs in serum: novel biomarkers for patients with bladder cancer? World J Urol. 32:353–358. 2014. View Article : Google Scholar

32 

Zhu HT, Dong QZ, Wang G, Zhou HJ, Ren N, Jia HL, Ye QH and Qin LX: Identification of suitable reference genes for qRT-PCR analysis of circulating microRNAs in hepatitis B virus-infected patients. Mol Biotechnol. 50:49–56. 2012. View Article : Google Scholar

33 

Lardizábal MN, Nocito AL, Daniele SM, Ornella LA, Palatnik JF and Veggi LM: Reference genes for real-time PCR quantification of microRNAs and messenger RNAs in rat models of hepatotoxicity. PLoS ONE. 7:e363232012. View Article : Google Scholar : PubMed/NCBI

34 

Song J, Bai Z, Han W, Zhang J, Meng H, Bi J, Ma X, Han S and Zhang Z: Identification of suitable reference genes for qPCR analysis of serum microRNA in gastric cancer patients. Dig Dis Sci. 57:897–904. 2012. View Article : Google Scholar

35 

Kreth S, Heyn J, Grau S, Kretzschmar HA, Egensperger R and Kreth FW: Identification of valid endogenous control genes for determining gene expression in human glioma. Neurooncol. 12:570–579. 2010.

36 

Hu Z, Dong J, Wang LE, et al: Serum microRNA profiling and breast cancer risk: The use of miR-484/191 as endogenous controls. Carcinogenesis. 33:828–834. 2012. View Article : Google Scholar : PubMed/NCBI

37 

Peltier HJ and Latham GJ: Normalization of microRNA expression levels in quantitative RT-PCR assays: Identification of suitable reference RNA targets in normal and cancerous human solid tissues. RNA. 14:844–852. 2008. View Article : Google Scholar : PubMed/NCBI

38 

Wang Y, Tang N, Hui T, Wang S, Zeng X, Li H and Ma J: Identification of endogenous reference genes for RT-qPCR analysis of plasma microRNAs levels in rats with acetaminophen-induced hepatotoxicity. J Appl Toxicol. 33:1330–1336. 2013.PubMed/NCBI

39 

Andersen CL, Jensen JL and Ørntoft TF: Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 64:5245–5250. 2004. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Wang L, Liu Y, Du L, Li J, Jiang X, Zheng G, Qu A, Wang H, Wang L, Zhang X, Zhang X, et al: Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer. Mol Med Rep 12: 615-622, 2015.
APA
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G. ... Wang, C. (2015). Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer. Molecular Medicine Reports, 12, 615-622. https://doi.org/10.3892/mmr.2015.3428
MLA
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G., Qu, A., Wang, H., Wang, L., Zhang, X., Liu, H., Pan, H., Yang, Y., Wang, C."Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer". Molecular Medicine Reports 12.1 (2015): 615-622.
Chicago
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G., Qu, A., Wang, H., Wang, L., Zhang, X., Liu, H., Pan, H., Yang, Y., Wang, C."Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer". Molecular Medicine Reports 12, no. 1 (2015): 615-622. https://doi.org/10.3892/mmr.2015.3428
Copy and paste a formatted citation
x
Spandidos Publications style
Wang L, Liu Y, Du L, Li J, Jiang X, Zheng G, Qu A, Wang H, Wang L, Zhang X, Zhang X, et al: Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer. Mol Med Rep 12: 615-622, 2015.
APA
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G. ... Wang, C. (2015). Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer. Molecular Medicine Reports, 12, 615-622. https://doi.org/10.3892/mmr.2015.3428
MLA
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G., Qu, A., Wang, H., Wang, L., Zhang, X., Liu, H., Pan, H., Yang, Y., Wang, C."Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer". Molecular Medicine Reports 12.1 (2015): 615-622.
Chicago
Wang, L., Liu, Y., Du, L., Li, J., Jiang, X., Zheng, G., Qu, A., Wang, H., Wang, L., Zhang, X., Liu, H., Pan, H., Yang, Y., Wang, C."Identification and validation of reference genes for the detection of serum microRNAs by reverse transcription‑quantitative polymerase chain reaction in patients with bladder cancer". Molecular Medicine Reports 12, no. 1 (2015): 615-622. https://doi.org/10.3892/mmr.2015.3428
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